Light and electron microscopic study on the endocrine cells of the pancreas in a marine teleost,Fugu rubripes rubripes

Summary The pancreatic endocrine tissue of Fugu rubripes rubripes consists of numerous round principal islets (Brockmann bodies) of various sizes scattered around the gall-bladder. The endocrine cells are divided into A-, B-, D-, and F_f-cells. Each cell type was identified by comparing thick and thin sections in both light and electron microscopy. Aldehyde-fuchsin positive B-cells contain numerous round secretory granules (average diameter 300 nm) each of which has a round compact core of moderate density; a narrow space exists between this core and the limiting membrane. Grimelius' silver positive A cells contain round secretory granules (average diameter 360 nm) with a hexagonal or tetragonal crystalline core (average diameter 170 nm) of high density; the silver grains preferentially appear in the space between the limiting membrane and the core. The crystalline core of each -granule often contains an appendix-like structure of variable shape. D cells blackened by the silver impregnation method of Hellman and Hellerström (1960) have round secretory granules (average diameter 320 nm) filled with a flocculent material of low density. The fourth cell type (F_f-cell) has a clear cytoplasm after differential staining for light microscopy. By electron microscopy, this cell has elongated fusiform secretory granules (520 nm average length × 230 nm average width) filled with numerous filaments arranged in parallel with the longitudinal axis. Figures suggesting granule formation in the sacs of the Golgi apparatus were obtained in all of islet cell types. Equivalents of emiocytotic release of secretory granules were encountered in the A and F_f cells.     

Correction to: Rhamphocottus nagaakii (Cottoidea: Rhamphocottidae), a new species of grunt sculpin from the northwestern Pacific,with notes on the phylogeography of the genus Rhamphocottus

A new species of grunt sculpin, Rhamphocottus nagaakii inhabiting the northwestern Pacific, previously identified as Rhamphocottus richardsonii Günther 1874, is described based on genetic evidence and morphological differences. The new species can be distinguished based on morphometric characters related to the head, including head length [45.3–54.6% of standard length (SL)], postorbital head length (18.8–25.5% SL) and the length of pectoral-fin base (15.8–20.7% SL), which are smaller than in R. richardsonii (53.6–60.5% SL, 26.2–31.7% SL, and 19.5–25.2% SL, respectively). Genetic differences between two species markedly exceed levels for intra-specific differences. Rhamphocottus nagaakii is considered to have arisen from a common ancestor of the two species, which probably inhabited somewhere the North Pacific Rim around the Aleutian Archipelago. During a period of cooling in the Pliocene or the Miocene, R. nagaakii and R. richardsonii became separated to the southern regions of the northwestern and northeastern Pacific, and subsequently underwent speciation.

     

Discovery of atrial natriuretic factor in the brain: Its characterization and cardiovascular implication

1. We have devised a radioimmunoassay for atrial natriueretic factor (ANF). Its application to rat brain extract led to the discovery of ANF in the brain. In addition to the hypothalamus and the pontine medullary region, it was widely distributed. 2. ANF in the brain is stored in a low molecular weight form, in contrast to pro-ANF in the atria. Thus, the processing of pro-ANF in the bran neuronal cells is different from that in the atria. 3. ANF was found in the anterior and posterior lobes of the pituitary, the peripheral ganglia, adrenergic neurons, and the adrenal medulla. 4. Brain ANF suppressed stimulated dipsogenesis, basal and stimulated vasopressin release, and angiotensin II-stimulated pressor effects. 5. ANF in the peripheral neuronal system inhibits catecholamine synthesis and release. Thus, central ANF functions to reduce the peripheral fluid volume and vascular tone in concert with the peripheral ANF.     

Weather Effects on the Patterns of People's Everyday Activities: A Study Using GPS Traces of Mobile Phone Users

This study explores the effects that the weather has on people''s everyday activity patterns. Temperature, rainfall, and wind speed were used as weather parameters. People''s daily activity patterns were inferred, such as place visited, the time this took place, the duration of the visit, based on the GPS location traces of their mobile phones overlaid upon Yellow Pages information. Our analysis of 31,855 mobile phone users allowed us to infer that people were more likely to stay longer at eateries or food outlets, and (to a lesser degree) at retail or shopping areas when the weather is very cold or when conditions are calm (non-windy). When compared to people''s regular activity patterns, certain weather conditions affected people''s movements and activities noticeably at different times of the day. On cold days, people''s activities were found to be more diverse especially after 10AM, showing greatest variations between 2PM and 6PM. A similar trend is observed between 10AM and midnight on rainy days, with people''s activities found to be most diverse on days with heaviest rainfalls or on days when the wind speed was stronger than 4 km/h, especially between 10AM–1AM. Finally, we observed that different geographical areas of a large metropolis were impacted differently by the weather. Using data of urban infrastructure to characterize areas, we found strong correlations between weather conditions upon people''s accessibility to trains. This study sheds new light on the influence of weather conditions on human behavior, in particular the choice of daily activities and how mobile phone data can be used to investigate the influence of environmental factors on urban dynamics.     

Simultaneous determination of dibucaine and naphazoline in human serum by monolithic silica spin column extraction and liquid chromatography-mass spectrometry

A simple, sensitive, and specific liquid chromatography-mass spectrometry (LC-MS) method for simultaneous determination of dibucaine and naphazoline from serum was developed and validated. The extraction procedure was performed using a monolithic silica spin column. Chromatographic separation of dibucaine and naphazoline was achieved on a C(18) reverse phase column with a mobile phase gradient (mobile phase A: 10mM ammonium formate and mobile phase B: acetonitrile) at a flow rate of 0.2mL/min. LC-MS was operated under the selective ion monitoring mode using the electrospray ionization technique in the positive mode. The retention times for naphazoline, dibucaine, and the internal standard (IS) were 6.7, 7.8, and 8.0min, respectively. A linear graph was obtained for dibucaine and naphazoline with correlation coefficients >0.998 for all analytes by this method. The limit of quantification of dibucaine and naphazoline was 10 and 25ng/mL, respectively. The mean recoveries were greater than 70%. Both compounds were stable under conditions of short-term storage, long-term storage as well as after freeze-thaw cycles. Monolithic spin column extraction and LC-MS analysis enabled the separation of dibucaine and naphazoline within 20min.     

Development of novel whole-cell immunoadsorbents by yeast surface display of the IgG-binding domain

The ZZ domain derived from Staphylococcus aureus, which binds to the Fc part of immunoglobulin G (IgG), was displayed on the cell surfaces of yeast Saccharomyces cerevisiae by cell-surface engineering using the C-terminal half of alpha-agglutinin under control of the 5'-upstream region of the isocitrate lyase gene from Candida tropicalis (UPR-ICL). Display of ZZ on the cell surface was confirmed by immunofluorescence microscopy. Enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA using the S. cerevisiae cells displaying ZZ detected IgG and antigen (human serum albumin) down to a concentration of 1-10 ng/ml in both cases. The detection range covered by these assay systems was wide and could be varied by adjusting the amount of cells and reaction times with horseradish peroxidase (HRP) substrate. Moreover, yeast cells displaying ZZ were successfully used for repeated affinity purification of IgG from serum. These results indicate that S. cerevisiae displaying ZZ may constitute novel and genetically renewable whole-cell immunoadsorbents widely applicable to immunoassays and affinity purification.     

Direct transformation of the liverwort <Emphasis Type="Italic">Marchantia polymorpha</Emphasis> L. by particle bombardment using immature thalli developing from spores

The liverwort, Marchantia polymorpha L., belongs to a group of basal land plants and is an emerging model for plant biology. We established a procedure to prepare sporangia of M. polymorpha under laboratory conditions by promoting its transition to reproductive development by far-red light irradiation. Here we report an improved direct transformation system of M. polymorpha using immature thalli developing from spores. Hygromycin-resistant transformants were obtained on selective media by transformation with a plasmid carrying the hygromycin-phosphotransferase gene (hpt) conferring hygromycin resistance in 4 weeks. The aminoglycoside-3'-adenyltransferase gene (aadA) conferring spectinomycin resistance was also successfully used as an additional selectable marker for nuclear transformation of M. polymorpha. The availability of the aadA gene in addition to the hpt gene should make M. polymorpha a versatile host for genetic manipulation. DNA gel-blot analyses indicated that transformed thalli carried a variable number of copies of the transgene integrated into the genome. Although the previous system using thalli grown from gemmae required a two-step selection in liquid and solid media for 8 weeks, the system reported here using thalli developing from spores allows generation of transformants in half the time by direct selection on solid media, facilitating genetic analyses in this model plant.     

Stabilization mechanism of cytochrome c552 from a moderately thermophilic bacterium, Hydrogenophilus thermoluteolus

Cytochrome c(552) (PH c(552)) from moderately thermophilic Hydrogenophilus thermoluteolus exhibits stability intermediate between those of cytochrome c(552) (HT c(552)) from thermophilic Hydrogenobacter thermophilus and cytochrome c(551) (PA c(551)) from mesophilic Pseudomonas aeruginosa. To understand the mechanism of stabilization of PH c(552), we introduced mutations into PH c(552) at five sites, which, in HT c(552), are occupied by the amino acids responsible for stability higher than the less stable PA c(551). When PH c(552) Val-78 was mutated to Ile, as found in HT c(552), the resulting variant showed increased stability. Mutation of Ala-7, Met-13, and Tyr-34 to the corresponding residues in PA c(551) (Phe, Val, and Phe, respectively) resulted in destabilization. We also found that PH c(552) Lys-43 contributed to stability through the formation of an attractive electrostatic interaction with Asp-39. These results suggest that the intermediate stability of PH c(552) is due to the amino acids at these five sites.     

Early development of the Japanese spiny oyster (Saccostrea kegaki): characterization of some genetic markers

The phylum Mollusca is one of the major groups of Lophotrochozoa. Although mollusks exhibit great morphological diversity, only a few comparative embryological studies have been performed on this group. In the present study, to begin understanding the molecular development of the diverse morphology among mollusks, we observed early embryogenesis in a bivalve, the Japanese spiny oyster, Saccostrea kegaki. Although several studies have begun to reveal the genetic machinery for early development in gastropods, very little molecular information is available on bivalve embryogenesis. Thus, as a step toward identifying tissue-specific gene markers, we sequenced about 100 cDNA clones picked randomly from a gastrula-stage cDNA library. This basic information on bivalve embryology will be useful for further studies on the development and evolution of mollusks.